This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is part of a Roadmap Consortium (U54 RR02437, T. Woodruff, PI) entitled "The Oncofertility Consortium: Fertility Preservation for Women". The goal is to provide leadership in state-of-the-art cryopreservation technology to practitioners who are preserving human ovarian tissues under the aegis of the National Physicians Cooperative. The specific aim is to optimize ovarian cryopreservation methods in nonhuman primates for restoring fertility in female cancer survivors by comparing novel vitrification freeze-thaw methods to the currently utilized slow freeze method. Although ovarian transplantation after cryopreservation has yielded live human offspring, autologous transfer in many cancer patients carries the risk of re-seeding the patient with her own cancer. In these patients, encapsulated 3-dimensional (3D) culture of isolated secondary follicles post-thaw may be the best option for fertility preservation. Since slow freezing damages secondary follicles, optimization of ovarian tissue vitrification could bridge a technical gap for patients to utilize frozen ovarian tissue outside of transplantation. For the first time, macaque follicles isolated from vitrified tissue can survive, grow, form an antrum and produce steroid hormones, indicating functional preservation of cryopreserved secondary follicles. The addition of non-permeating polymers to the vitrification solution improved tissue morphology and follicle function in 3D culture. Current efforts are aimed at producing a mature oocyte derived from 3D follicle culture that can fertilize, undergo embryonic development and ultimately produce a live offspring. This project is facilitated by interactions with the Biomaterials Core at Northwestern University. Expected advances will be transferred to clinical efforts at Fertility Centers.